Objective To analyze the effectiveness and safety of bisphosphonates in the treatment of pa-tients with calcification defense.Methods PubMed,Embase databases,CNKI and Wanfang were searched to collect the case reports and clinical studies of bisphosphonates for calcification defense.Then,the relevant infor-mation of patients was extracted for statistical analysis.Results A total of 18 case reports were selected involving 20 patients.Thirteen patients(65.0％)were treated with pamidronate,four(20.0％)were treated with etidr-onate,two(10.0％)were treated with alendronate,and one(5.0％)was treated with zoledronic acid.Thirteen patients(65.0％)recovered completely,the recovery time of whom ranged from half month to nine months.The tolerance of bisphosphonates in most patients(90.0％)was good,while one patient who did not tolerate pamidr-onate recovered after the frequency of administration was adjusted and one patient with high dosage of etidronate returned to normal after the discontinuation of the usage.Conclusions Bisphosphonates,an inhibitor of bone resorption,is effective and safe in the treatment of patients with calcification defense.

Objective:Human leukocyte antigen(HLA)B27 is a susceptibility allele of ankylosing spondylitis(AS),and HLA-B27 antigen typing is an important indicator for clinical diagnosis of AS,but current typing methods such as sequence specific primer polymerase chain reaction(PCR-SSP)still possess limitation.Therefore,this study aims to analyze the correlation between B27 subtypes and susceptibility to AS in Hunan Province by applying high-resolution polymerase chain reaction-sequence-based typing(PCR-SBT). Methods:Peripheral blood of 116 patients with suspected AS(suspected AS group)and 121 healthy volunteers(control group)admitted to the Second Xiangya Hospital from January 2020 to December 2020 were collected for HLA-B genotyping by PCR-SBT.Among the patients in the suspected AS group,23 patients were finally diagnosed with AS(confirmed AS group),and the remaining 93 undiagnosed patients served as the non-confirmed AS group.PCR-SBT and PCR-SSP were used to detect HLA-B27 typing in 116 patients with suspected AS,and the results of the 2 methods were compared. Results:The HLA-B27 allele frequency in the suspected AS group was significantly higher than that in the control group[11.63%vs 2.48%;P<0.001,odds ratio(OR)=5.18,95%confidence interval(CI)2.097 to 12.795].B*27:04,B*27:05,B*27:06,and B*27:07 were detected in the suspected AS group and the control group.The frequency of the B*27:04 allele in the suspected AS group was significantly higher than that in the control group(9.48%vs 1.24%;P<0.001,OR=8.346,95%CI 2.463 to 28.282).The positive rate of B27 in the suspected AS group and the confirmed AS group(B27+/+ and B27+/-)was significantly higher than that in the control group(χ2=16.579,P<0.001;χ2=94.582,P<0.001,respectively).Among the confirmed AS group,21 were HLA-B27 carriers,and the B27 positive rate in the confirmed AS group was 91.3%.PCR-SBT could achieve high resolution typing of the HLA-B gene locus,with higher sensitivity,specificity,positive predictive value,negative predictive value,and accuracy than PCR-SSP. Conclusion:PCR-SBT typing analysis shows a strong correlation between HLA-B * 27:04 and AS in Hunan province.The PCR-SBT method can be used as the preferred option for the auxiliary diagnosis of clinical AS.

Objective:To investigate the advantages and disadvantages and clinical effects of integrated acetabular quadrilateral surface buttress plate in the treatment of acetabular both column fractures.Methods:From September 2017 to March 2019, 14 patients with acetabular both column fractures were treated with integrated quadrilateral acetabular buttress plate and were followed up. The clinical data were retrospectively analyzed. There were 10 males and 4 females with an average age of 53.2 years (range, 26-75 years). Fracture classification were both column fractures according to the Letournel-Judet classification system, including 11 cases of combined posterior wall without posterior dislocation. The time from injury to operation was 8-19 days, with an average of 10.3 days. All operations were performed with a supra-ilioinguinal approach. After satisfactory reduction, the integrated acetabular quadrilateral surface buttress plate was used to fix the anterior and posterior column and quadrilateral fractures at the same time. After operation, Matta criteria was used to evaluate the quality of fracture reduction on postoperative images, and modified Merle d'Aubigné-Postel score to evaluate hip joint function.Results:The average operation time was 180 min (range, 120-320 min), and the intraoperative average blood loss was 980 ml (range, 700-1 600 ml). Operations were performed successfully with none intraoperative reshaping conducted, and the plate could fit into the bone surface well. All 14 patients were followed up with an average of 15.6 months (range, 9-20 months). During the follow-up, all fractures healed with an average of 3 months (range, 2.5-5 months). The reduction quality evaluated by the Matta criteria were 7 anatomical, 5 satisfactory, and 2 unsatisfactory, with an excellent and good rate of 85.7% (12/14). The hip joint function at the last follow-up was in accordance with the modified Merle d'Aubigné-Postel score was 12-18 points, with an average of 16.9 points, of which 8 excellent, 4 good, and 2 fair. The excellent and good function rate was 85.7% (12/14). During the operation, 1 patient had a rupture of about 1 cm of the peritoneum during the separation of the spermatic cord, which was immediately sutured and repaired. One patient had obturator nerve paralysis after the operation, without special treatment, and the paralysis symptoms were relieved during the 6th-month follow-up. There were no other operation-related complications.Conclusion:For both-column fractures characterized by central dislocation of the femoral head, the integrated acetabular quadrilateral buttress plate can simultaneously fix the anterior and posterior columns and quadrilateral surface at the same time. Satisfactory reduction and functional outcomes could be achieved by the novel plates.

Objective:To introduce a novel anatomical supra-pectineal quadrilateral surface buttress plate, and explore its advantages, disadvantages and clinical efficacy in the treatment of complex acetabular fractures.Methods:Data of eighteen cases of acetabular fractures treated in our department from March 2017 to March 2019were retrospectively analyzed. There were 15 males and 3 females with an average age of 49.6 years (range, 27-62 years). According to the Letournel-Judet’s classification, there were 3 cases of anterior column fractures (involving quadrilateral surface), 2 cases of "T" shaped, 8 of cases anterior column and posterior hemi-transverse (ACPHT) and 5 cases of double columns fractures. The time from injury to operation was 5-14 days (mean, 7 days). Before operation, all patients were taken pelvic anterior-posterior X-ray and CT scan. All patients were adopted supra-ilioinguinal approach and fixed by supra-pectineal quadrilateral surface buttress plate. Surgical time, intraoperative blood loss and postoperative complications were collected. After operations all patients were taken pelvic X-ray in anterior-posterior and Judet position, as well as CT scan. The radiographic quality of reduction was evaluated by Matta criteria and hip functions were evaluated by Matta Modified Merle d’Aubigné-Postel scoring system.Results:The average surgical time was 130 min (range, 100-200 min). The average blood loss was 560 ml (range, 400-900 ml). An additional shaping was unnecessary in 18 patients who were implanted with the anatomical supra-pectineal quadrilateral surface buttress plate and all patients were operated successfully. All patients were followed up for an average time of 9 months (range, 6-15 months). The quality of fracture reduction was evaluated according to Matta standard, of which there were 11 cases excellent, 4 cases good, and 3 cases poor, with an excellent rate of 83.3% (15/18). At the latest follow-up, the function of the hip joint was evaluated according to the Matta Modified Merle d’Aubigné-Postel scoring system for a mean of 15.8 scores (range, 12-18 scores), of which there were 10 cases excellent, 6 cases good, 2 cases fair, with an excellent rate of 88.9% (16/18). All patients recovered well, and there were 2 cases of lateral femoral cutaneous nerve injury postoperative who had recovered one month later. There was no other postoperative complications.Conclusion:The anterior and posterior column and quadrilateral surface could be stabilized simultaneously by the anatomical supra-pectineal quadrilateral surface buttress plate without intraoperative shaping. Moreover, it is more likely to obtain a satisfactory clinical outcome by using this special plate with lower complications and rigid fixation. Therefore, it is a kind of internal fixation method worthy of promotion in the treatment of complex acetabular fractures.

OBJECTIVE: To study the effect of aquaporin 4(AQP4) in regulating the permeability of blood-brain barrier(BBB) induced by subacute 1,2-dichloroethane(1,2-DCE) inhalation. METHODS: Specific pathogen free healthy CD-1 male Aqp4 genetically engineered mice(Aqp4~(+/+)and Aqp4~(-/-)) were randomly divided into control and low-, medium-and high-dose groups. The mice were exposed to 1,2-DCE at the dosages of 0.00, 100.00, 350.00 and 700.00 mg/m~3 for 6 hours per day for consecutive 28 days by systemic dynamic inhalation. After the end of 1,2-DCE exposure, the BBB permeability was evaluated by Evans blue staining. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of genes related to BBB tight junction protein(Tjp)1, Tjp2, Tjp3, claudin(Cldn)3, Cldn5, Cldn11, occludin(Ocln), matrix metalloproteinase(Mmp)2, Mmp9 and Na-K-Cl cotransporter-1(Nkcc1). RESULTS: The BBB permeability in mice showed significant change with 1,2-DCE dose and Aqp4 genotype(P<0.01). The BBB permeability of Aqp4~(+/+) genotype mice was higher in low-, medium-and high-dose groups than that of control group(all P values were <0.05). The permeability of BBB was lower in Aqp4~(+/+) genotype mice in the control group than that of Aqp4~(-/-) genotype mice in the same group(P<0.05), but BBB permeability was higher in Aqp4~(+/+) genotype mice in the three dose groups than that of Aqp4~(-/-) genotype mice in the same group(all P values were <0.05). The Cldn3 and Olcn mRNA relative expression in the brain cortex had statistical difference in mice with different genotype(all P values were <0.01). The mRNA relative expressions of Cldn3 and Olcn in the brain cortex were higher in Aqp4~(-/-) genotype mice than that of Aqp4~(+/+) genotype mice(all P values were <0.01). The relative mRNA expression levels of Tjp1, Tjp2, Tjp3, Cldn5, Cldn11, Mmp2, Mmp9 and Nkcc1 in the cerebral cortex of mice were not statistically significant in aspect of 1,2-DCE exposure dose and genotype(all P values were >0.05). CONCLUSION: Exposure to 1,2-DCE can increase BBB permeability in mice, and the mechanism may be associated with 1,2-DCE-induced down-regulation of Aqp4 and up-regulation of mRNA expression of the cerebral cortex TJP-related molecules Cldn3 and Ocln.

OBJECTIVE: To investigate the effect of 1,2-dichloroethane(1,2-DCE) acute inhalation exposure on the differential gene expression of phase Ⅰ metabolic enzymes. METHODS: The specific pathogen free SD rats were randomly divided into control group(16 rats), low-and high-dose groups(24 rats in each group, half males and half females). Low-and high-dose group were given daily 600, 1 800 mg/m~(3 ) of 1,2-DCE, and the control group given the fresh air by dynamic inhalation for 8 hours per day for consecutive 7 days. After the end of exposure, the relative mRNA expression of cytochrome P450 2 E1(CYP2 E1), alcohol dehydrogenase(ADH1) and acetaldehyde dehydrogenase 3 alpha 1(ALDH3α1) in the liver tissue was detected by real-time fluorescence quantitative polymerase chain reaction. RESULTS: The relative expression of CYP2 E1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ADH1 in male low-and high-dose groups was higher than that in male control group(P<0.05). The relative expression of ADH1 in male high-dose group was higher than that in male low-dose group and female high-dose group(P<0.05). The relative expression of ALDH3α1 in high-dose group was higher than that in control group and low-dose group(P<0.05). CONCLUSION: High dose 1,2-DCE could increase the gene expression of phase Ⅰ metabolic enzymes in rat liver. The 1,2-DCE has more obvious effect in male rats than in female rats.

OBJECTIVE: To investigate the effects of subacute systemic inhalation exposure of 1,2-dichloroethane(1,2-DCE) on learning and memory in NIH mice. METHODS: Forty-five specific pathogen free healthy 7-week-old NIH mice were randomly divided into control,low-dose and high-dose groups with 5 female mice and 10 male mice in each group. The mice were exposed to 1,2-DCE at dosages of 0. 00,100. 00 and 350. 00 mg/m3 for 6 hours per day for consecutive 28 days by dynamic systemic inhalation. The neurobehavioral tests of mice were performed before and after the first to fourth weeks of exposure using the Morris water maze test. RESULTS: There was no significant difference in body weight and swimming speed among the three groups of mice( P > 0. 05). The navigation experiment results showed that the escape latency of mice in both low-and high-dose groups were longer than that of the control group at the same time point(P < 0. 05) during 1-4 weeks after exposure. In the control group,the escape latency was shorter than that of the same group before exposure( P < 0. 05). The escape latency of high-dose group prolonged with the increase of exposure time,and in the 4 th week the escape latency was significantly higher than that of the same group before exposure( P < 0. 05).The experiment results of space exploration indicated that the first time of crossing platform in low-and high-dose groups were longer than that of the control group at the second to the fourth week( P < 0. 05). The target quadrant retention time and the number of crossing the platform in the low-and high-dose groups were lower than those in the control group( P <0. 05). CONCLUSION: Subacute inhalation exposure of 1,2-DCE can impair the learning and memory ability of NIH mice.The high-dose exposure may reduce learning ability in mice in a time-effect manner.

OBJECTIVE: To compare the effects of different anesthetics and blood sampling methods on blood routine test results in experimental animals. METHODS: A total of 42 specific pathogen free( SPF) male Sprague Dawley( SD) rats and 59 SPF male Kunming( KM) mice were randomly divided into 4 groups( control group,ether group,chloral hydrate group and pentobarbital sodium group). Ether group animals were treated with ether inhalation anesthesia; animals in chloral hydrate group and pentobarbital sodium group were injected intraperitoneally with chloral hydrate or pentobarbital sodium. The control group received no anesthesia treatment. Blood samples were collected by different ways: orbital venous plexus,abdominal aorta or eyeball enucleation. White blood cell( WBC) count,red blood cell( RBC) count,platelet(PLT) count,hemoglobin(Hb) level and hematocrit(HCT) in blood samples were analyzed. RESULTS: The RBC count,Hb level and HCT of SD rats in pentobarbital sodium group were significantly lower than those in control group( P <0. 05). The HCT of SD rats in ether group was lower than that in control group( P < 0. 05). The WBC count of orbital venous plexus of KM mice was lower than that taken by eyeball enucleation in control group( P < 0. 05),but the WBC count of orbital venous plexus was higher than that taken by eyeball enucleation in chloral hydrate group( P < 0. 05). The RBC count,Hb level,HCT of KM mice in pentobarbital sodium group were significantly lower than those in control group(P < 0. 05). CONCLUSION: The anesthetic can affect the blood routine test results of experimental animals. Different blood sampling methods have effects on blood routine test results of KM mice.

OBJECTIVE: To predict the sensitizing potency and optimal sensitization dose of trichloroethylene( TCE) by an in vitro skin sensitization test on a human acute monocytosis cell line( THP-1).METHODS: THP-1 cells were cultured in vitro and exposed to 2,4-dinitrochlorobenzene( DNCB),sodium dodecyl sulfate( SDS),tert-butylhydroquinone( tBHQ)and TCE for 24 hours.Flow cytometry was used to detect the expression of cell surface marker such as cluster of differentiation( CD) 86 and CD54,and the optimal dose range for sensitization detection was determined.With the relative fluorescence intensity( RFI),CD86 ≥ 150 and CD54 ≥ 200 as the standard,the sensitizing potency and optimal sensitization dose of TCE were predicted.RESULTS: The concentration range of reagents for sensitization test on THP-1 cells was the dose range at which the relative cell survival rate reached 75.0%-100.0%.DNCB at the doses of 20.83,25.00 and 30.00 μmol/L,tBHQ at the dose of 5.80 μmol/L,TCE at the doses of 8.33,10.00 and 12.00 mmol/L,can cause sensitivity.SDS was recognized as a negative sensitizer.The expression of CD86 and CD54 was the highest when the concentration of TCE was 8.33 mmol/L,which was considered as the best sensitization dose.CONCLUSION: The optimum sensitization dose of TCE is 8.33 mmol/L,which can provide the basis for dose design in future study of TCE sensitization pathways.

OBJECTIVE: To explore the molecular mechanism underlying 1,2-dichloroethane(1,2-DCE) induced apoptosis by screening differentially expressed proteins in human astrocytes( HAs). METHODS: HAs were cultured in complete medium with 1,2-DCE at various concentrations of 0-80 or 0-40 mmol/L. After 24 hours,apoptosis of HAs was evaluated using flow cytometry and staining with annexin Ⅴ-fluoresce in isothiocyanate and propidium iodide. An AAH-APO-1-2 protein chip was used to screen differentially expressed proteins and quantitative real-time polymease chain reaction(qRT-PCR) was used to verify related differentially expressed genes(DEGs). RESULTS: At 1,2-DCE concentrations of0-80 mmol/L,the total apoptosis rate of HAs increased with 1,2-DCE concentrations in a dose-dependent manner( P <0. 01). Seven different kinds of proteins were screened out by apoptotic protein chip. Among them,the expression of insulin-like growth factor-binding protein( IGFBP)-1,IGFBP-4 and cytochrome C( Cyto C) were up-regulated,while the expression of P27,cysteine aspartic acid specific protease-3( Caspase-3),B-cell lymphoma-2 interacting mediator of cell death( BIM) and BH3 interacting domain death agonist( BID) were down-regulated compared with the control group. The result of DEGs verified by qRT-PCR showed that the expression of mRNA of IGFBP-1,IGFBP-4 and Cyto C at 1,2-DCE concentrations of 40 mmol/L was up-regulated. This result was in consistent with the trend of target expression in the protein chip. The mRNA expression of Caspase-3,BIM and BID was also up-regulated. CONCLUSION: 1,2-DCE induces apoptosis of HAs through mitochondrial pathway.